Fecal pollution can degrade water quality and restrict its use for drinking, recreational activities, plant irrigation, and for harvesting sea foods. Understanding the source(s) of pollution in bodies of water is paramount to assessing health risk and implementing remedial actions. Currently, water quality managers do not have simple and accurate tools to determine the specific sources of pollution. We have reported a ribotyping test that discriminates human and nonhuman sources of E. coli, but is a two-week procedure that is expensive and not practical for broad application to other laboratories. We are confident that these problems can be remedied by extending our previous research to produce a simple PCR-based test that many laboratories can perform. To determine the feasibility of this approach, we propose to use Amplified Fragment Length Polymorphism (AFLP) and subtractive hybridization techniques to determine DNA sequences that differentiate human and non-human strains of E. coli. If Phase I proves successful, then we will be able to expand and refine the research leading to a one to two-day test. The simpler test format will promote commercialization by making it much more feasible for various water quality laboratories to perform.
Anticipated Results/Potential Commercial Applications of Research: Phase I will determine whether there are DNA sequences in E. coli that can discriminate as either human or non-human. If both Phases are successful, the nation will have important tools for resolving issues of water quality. The envisioned test can be used bu numerous laboratories, thereby expanding application to many more situations than presently possible. Government regulatory agencies will likely see the greatest benefit through commercialized tests of water used for human consumption and recreation. The cost/benefit ratio will be very low due to the potential benefits in reducing the incidence of human waterborne disease.
